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AIMS This study aimed to evaluate the effectiveness of selected essential oils (EOs) and hydrolates (Hs) against L. monocytogenes biofilms on polystyrene (PS) and stainless steel (SS) surfaces. METHODS AND RESULTS Among others, Origanum hirtum EO, Corydothymus capitatus EO and Citrus aurantium H were selected to treat L. monocytogenes biofilms during and after biofilm formation. Sub-Minimum Inhibitory Concentrations (MICs) of C. capitatus EO (0.31 µL/mL) showed the highest inhibiting effect against biofilm formation on PS, while on SS no significant differences between the EOs were observed (43.7%-88.7% inhibition). Overall, the tested biosanitizers showed limited activity as biofilm removal agents. Although generally less effective, C. aurantium H exhibited good biofilm inhibition performance at 62.5 µL/mL, particularly on PS. Confocal Laser Scanning Microscopy proved that sub-MICs of the biosanitizers drastically changed L. monocytogenes biofilm architecture, with bacterial cells elongation in the presence of C. capitatus EO. CONCLUSIONS Our findings suggest that the tested EOs and H are able to control Listeria biofilms, particularly preventing biofilm formation on both materials. Considering its mild aroma and hydrophilicity, the H exhibited promising perspectives of application. SIGNIFICANCE AND IMPACT OF STUDY This study raises the possibility of applying EOs and Hs to control biofilms on different surfaces in the food industry.